A High-Performance Liquid Chromatography Radio Method for Determination of l-Ascorbic Acid and Guanosine 5′-Diphosphate-l-Galactose, Key Metabolites of the Plant Vitamin C Pathway
✍ Scribed by Beata A. Wolucka; Mark W. Davey; Wout Boerjan
- Publisher
- Elsevier Science
- Year
- 2001
- Tongue
- English
- Weight
- 224 KB
- Volume
- 294
- Category
- Article
- ISSN
- 0003-2697
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✦ Synopsis
A simple, rapid, and quantitative high-pressure liquid chromatography radio method is described for the determination of in vivo (14)C-labeled l-ascorbate, dehydro-l-ascorbate, and total l-ascorbate of Arabidopsis thaliana cell suspensions upon incubation of cultures with exogenous d-[(14)C]mannose. The same radio-HPLC conditions can be used to follow the products of in vitro enzymatic conversions of GDP-d-mannose by enzyme extracts of A. thaliana, namely GDP-l-galactose, GDP-4"-keto,6"-deoxy-d-mannose, and GDP-l-fucose. In particular, an accurate assay for GDP-d-mannose 3",5"-epimerase, a key enzyme of the plant vitamin C pathway, is presented.